Expression and Characterization of the Bovine Histamine H1 Receptor in cDNA-Transfected C6 Astroglioma Cells

Rat C6 astroglioma cells (C6-bH1R cells) expressing cloned bovine histamine H1 receptors were established by transfection with a vector (pEF-BOS-bH1R) which carried a 2.7-kbp EcoRI fragment of the bovine H1 receptor cDNA [Yamashita, M. et al. (1991) Proc. Natl. Acad. Sci. USA 88, 11515–11519]. The c...

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Published in:Journal of biochemistry (Tokyo) 1994-06, Vol.115 (6), p.1155-1161
Main Authors: Mizuguchi, Hiroyuki, Ito, Seiji, Shevchenko, Valery I., Nagasawa, Yasuyuki, Yamashita, Masakatsu, Imamura, Ikuo, Horio, Yoshiyuki, Fujimoto, Katsumi, Fukui, Hiroyuki
Format: Article
Language:English
Subjects:
Animals
Astrocytes - metabolism
Astrocytoma - genetics
Astrocytoma - metabolism
astroglioma
bovine
Cattle
Cloning, Molecular
DNA, Complementary - genetics
expression
Genetic Vectors
histamine
histamine H1 receptor
Inositol Phosphates - metabolism
Radioligand Assay
Rats
Receptors, Histamine H1 - biosynthesis
Receptors, Histamine H1 - genetics
Recombinant Proteins - biosynthesis
Transfection - genetics
Tumor Cells, Cultured
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Summary:Rat C6 astroglioma cells (C6-bH1R cells) expressing cloned bovine histamine H1 receptors were established by transfection with a vector (pEF-BOS-bH1R) which carried a 2.7-kbp EcoRI fragment of the bovine H1 receptor cDNA [Yamashita, M. et al. (1991) Proc. Natl. Acad. Sci. USA 88, 11515–11519]. The cloned bovine H1 receptor in C6-bH1R cells was characterized by three established criteria: the [3H]mepyramine binding assay, the accumulation of inositol phosphates induced by histamine, and histamine-induced elevation of intracellular Ca2+ concentration ([Ca2+]1). The accumulation of inositol phosphates induced by histamine was time- and dose-dependent. The accumulation of inositol tris-phosphate was biphasic with a prompt increase to the maximal level, followed by a sustained submaximal level. The histamine-induced accumulation of inositol phosphates was suppressed by phorbol ester, but not by pertussis toxin. Results from the [3H]-mepyramine binding assay and histamine-induced elevation of [Ca2+], were characteristic of H1 receptors. Several compounds among tricyclic antidepressants, neuroleptics, and serotonin antagonists showed affinities to the cloned bovine H1 receptor with K1 values similar to reported values. Histamine neither induced cAMP accumulation nor attenuated forskolin-induced cAMP accumulation in C6-bH1R cells. C6-bH1R cells are particularly useful for studying the H1 receptor-mediated astroglial cell functions.
ISSN:0021-924X
DOI:10.1093/oxfordjournals.jbchem.a124472